Scaffolded fusion polypeptides and compositions and methods for making the same

Abstract

The present invention provides soluble forms of integral membrane proteins, or domains or portions thereof, that retain the biological activity of the integral membrane protein, domain or portion from which they are designed or derived and that can readily be expressed in high yield.

Description

1. FIELD OF THE INVENTIONThe present invention relates to novel compositions designed from integral membrane proteins that have improved solubility and retain biological activity.2. BACKGROUND OF THE INVENTIONMany important cellular and biological processes are mediated by integral membrane proteins. Often quantities of these integral membrane proteins are required to study their roles in the critical cellular and biological processes. However, integral membrane proteins are often difficult to obtain in quantity while retaining solubility and function.For example, according to one common technique, the integral membrane protein is extracted from the bilayer with a nonionic detergent. Unfortunately, the proteins recovered by such extraction techniques frequently lack activity.In another common technique, a soluble portion of the membrane protein, such as an extracellular domain, is expressed in the absence of the insoluble portions of the membrane protein (typically the transmembrane...

AI Technical Summary

Benefits of technology

"The present invention provides scaffolded fusion polypeptides that are designed to be more soluble and biologically active in a membrane-free and detergent-free aqueous environment. These scaffolded fusion polypeptides display improved solubility by fusing a scaffold domain to the functional domain of an integral membrane protein. The scaffold domain mimics the structure of the protein from which it is derived, and can be linked together to form polymeric scaffolded fusion polypeptides. The scaffolded fusion polypeptides are modular structures that can be designed and synthesized to span the membrane multiple times, allowing for the creation of soluble forms of membrane proteins."

Problems solved by technology

However, integral membrane proteins are often difficult to obtain in quantity while retaining solubility and function.

Unfortunately, the proteins recovered by such extraction techniques frequently lack activity.

However, this method is inadequate for expressing the soluble portions of many membrane proteins, especially those that traverse the membrane more than once.

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